| As the most common type of dementia,patients suffering from Alzheimer's disease(AD)is soaring worldwide.It has become a huge burden for the family and society.Nevertheless,the therapeutic effect of current therapies is unsatisfactory.Traditional Chinese medicine compound contains varieties of active ingredients,which character as acting on multiple targets and signaling pathways and exerting a holistic regulating effect.Guilingji is famous for its effect of tonifying kidney and anti-aging.Therefore,it is applicable to explore the role and mechanism of Guilingji in the treatment of AD.Objective:To evaluate the clinical effectiveness of Guilingji on the treatment of patients with Alzheimer's disease(AD),and effect of Guilingji on the learning and memory ability of SAMP8 mice and the expression of synapse and myelin associated protein,so as to provide scientific basis for the clinical application.Methods:1 Clinical research:60 AD patients with kidney deficiency and encephala reduction were randomized assigned to experimental group or control group,the experimental group accepted Guilingji for 24 weeks,the control group accepted Yinxingye tablet for 24weeks.At the baseline,the midpoint and endpoint,the changes of ADAS-cog,MMSE,ADL,TCM syndrome were observed.2 Experimental research:64 SPF grade the SAMP8 mice were randomly divided into four group:Model group,Guilingji low-dose(GLJ-L)group,Guilingji high-dose(GLJ-H)group and GinKgo Biloba extract(YXY)group.The SAMR1 mice was used as control(Control).Mice of GLJ-L?GLJ-H?YXY group were given the corresponding drugs.Mice of Control and Model group were given equal volume of distilled water.After 12 weeks of drug intervention,New object recognition experiment and Morris water maze experiment were used to assess the memory and learning ability of SAMP8 mice.The pathological morphology of brain tissue of SAMP8 mice were observed by HE and Nissl staining.Western blot was used to observed the expression of GAP43?SYP?MBP?BDNF?TrkB proteins in the hippocampus tissue.QPCR was used to detect the mRNA expression of these proteins.Results:1 Results of Clinical research:(1)Change in neuropsychological scale score:compared with baseline,the ADAS-cog scores of experimental group and control group were significantly decreased at the 12th week and 24th week(P<0.01 or P<0.05).There was no significant difference among the groups at the 12th week and endpoint(P>0.05).Compare with baseline,the MMSE scores of experimental group and control group were significantly increased at 12th week and 24th week(P<0.01).At the 12th week and endpoint,there was no statistical difference in MMSE score among groups(P>0.05).(2)Change in activity of daily living scale(ADL)score:no statistical difference was observed in experimental group and control group(P>0.05).(3)Change in TCM syndrome score:after 24 weeks of intervention,the TCM syndrome score of experimental group and control group was significantly decreased(P<0.01).In the experimental group,the effective rate of soreness and weakness of waist and knees,fatigue and sleepiness were significantly higher than that of control group(P<0.01),The total effective rate was 59.3%for experimental group,19.05%for control group.The effective rate was statistical difference among the two groups(P<0.01).(4)Safety evaluation:no severe adverse event was observed during the treatment period in experiment and control group.No abnormal change in blood routine,urine routine,liver function(AST,ALT),renal function(UREA,CRE)was observed.2.Results of Experimental research:(1)Effect of Guilingji on recognition memory ability of SAMP8 miceCompared with control group,the discrimination index of SAMP8 mice of model group showed a down-regulated trend,but the difference was not statistically significant(P>0.05).Compared with model group,the discrimination index of SAMP8 mice of GLJ-L,GLJ-H and YXY group showed an up-regulated trend,but the difference was not statistically significant(P>0.05).(2)Effect of Guilingji on spatial learning and memory of SAMP8 miceIn the orientation navigation experiment of Morris water maze,the escape latency of all mice in each group was gradually decreased with the training day.Compared with control group,the escape latency of model group was significantly longer(P<0.01).Compared with model group,the escape latency of GLJ-L,GLJ-H group and YXY group were significantly decreased(P<0.01).For the same group,compared with the escape latency of the first day,control group,GLJ-L,GLJ-H group showed a significant improvement in learning ability at the third day(P<0.05,P<0.05,P<0.01),however,the escape latency of GLJ-L,GLJ-H group were longer than control group.YXY group showed the tendency in the second day of training(P<0.05).During the same day,compared with control group,the escape latency of model group on the first day to the fifth day were longer(P<0.05,P<0.01).Compared with model group,the escape latency of GLJ-H group and YXY group were significant decreased from the second to the fifth day(P<0.01).GLJ-L group showed significant reduction on the fifth day(P<0.05).In the probe test:Compared with control group,the crossing times,percentage of swimming time and swimming distance in target quadrant of model group were significantly decreased(P<0.01,P<0.01,P<0.05).Compared with model group,crossing times of GLJ-L and GLJ-H were significantly increased(P<0.05).Percentage of swimming time in the target quadrant of GLJ-L,GLJ-H and YXY group were increased(P<0.05).Percentage of swimming distance in the target quadrant of GLJ-H was increased(P<0.01)(3)Effect of Guilingji on hippocampal pathological morphology of SAMP8 miceIn the control group,the neurons in CA1 area of hippocampus were orderly,tightly arranged,with regularly morphology,clearly boundary and nucleoli and numerous Nissl bodies.In the model group,Nissl bodies were decreased or even disappeared,which result in stained light.The scattered neurons were disordered distribution,with unclear boundary,karyopyknosis and even nuclei disappeared.Compared with the model group,neurons showed different degree of improvement in GLJ-L,GLJ-H and YXY group,such as the neuron tightly and neatly arranged with more cell layers and Nissl bodies within cytoplasm,which could be dark blue stained.(4)Effect of Guilingji on the expression of synapse and myelin associated proteins of SAMP8 miceCompared with control group,the expression of SYP mRNA and GAP43,MBP protein and mRNA in SAMP8 mice of model group were significantly decreased(P<0.01).The expression of protein and mRNA of GAP43,MBP were increased in GLJ-L and GLJ-H group(P<0.01 or P<0.05),as well as the same tendency of mRNA of SYP(P<0.01).(5)Effect of Guilingji on BDNF/TrkB signaling pathwayCompared with control group,the expression of protein and mRNA of BDNF were significantly reduced(P<0.01).But,the expression of protein of TrkB was not significantly reduced(P>0.05).Compared with model group,the expression of BDNF,TrkB protein were significantly increased in GLJ-H group(P<0.01,P<0.05),as well as the expression of mRNA of TrkB(P<0.01).Conclusions:1 Conclusion of clinical research:Guilingji could reduced score of ADAS-cog and TCM symptoms,and increased score of MMSE.Therefore,Guilingji could improve the cognitive function of patients with mild to moderate AD.No severe adverse event was observed during the trial.2 Conclusion of experimental research:Guilingji could improve learning and memory ability of SAMP8 mice,the mechanism may be related to reducing the lose of neurons in CA1 area,regulating the BDNF/TrkB signaling pathway and promoting the expression of synapse and myelin sheath associated protein. |
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